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Abstract:

A hydrolytic enzyme is to be stabilized in a liquid surfactant
preparation. This is achieved by using a component that stabilizes the
hydrolytic enzyme and encompasses a phthaloylglutamic acid and/or a
phthaloylaspartic acid.

2. The surfactant preparation according to claim 1, wherein the
phthaloylglutamic acid is contained in a quantity from 0.000001 to 10 wt
%, and the hydrolytic enzyme is contained in a quantity from
1.times.10.sup.-8 to 5 weight percent, based on active protein.

3. The surfactant preparation according to claim 1, wherein the
hydrolytic enzyme is selected from the group comprising a protease,
amylase, cellulase, glycosidase, hemicellulase, mannanase, xylanase,
xyloglucanase, xanthanase, pectinase, β-glucosidase, carrageenase,
lipase, and a mixture that encompasses at least two of said enzymes.

4. The surfactant preparation according to claim 1, wherein the
preparation is a washing agent, cleaning agent, or disinfection agent.

5. The surfactant preparation according to claim 1, wherein the
surfactant preparation moreover comprises at least one further ingredient
that is selected from the group consisting of builder, nonaqueous
solvent, acid, water-soluble salt, thickening agent, disinfecting
ingredient, and combinations thereof.

6. A washing or cleaning method, comprising: stabilizing a hydrolytic
enzyme in a washing using a component that stabilizes the hydrolytic
enzyme and comprises a phthaloylglutamic acid, wherein the enzyme is
selected from the group consisting of protease, amylase, cellulase,
glycosidase, hemicellulase, mannanase, xylanase, xyloglucanase,
xanthanase, pectinase, β-glucosidase, carrageenase, lipase, and
mixtures thereof.

Description:

FIELD OF THE INVENTION

[0001] The present invention generally relates to liquid enzyme-containing
surfactant preparations such as those utilized, for example, for washing,
cleaning, or disinfecting, and more particularly relates to a liquid
surfactant preparation of this kind in which a hydrolytic enzyme is
stabilized. The invention further relates to uses of enzyme stabilizers,
and to methods in which enzymes stabilized in this fashion are used. The
invention further relates to enzyme preparations stabilized in this
manner.

BACKGROUND OF THE INVENTION

[0002] Problems relating to the shelf stability of enzyme-containing
surfactant preparations, for example of washing, cleaning, or
disinfecting agents, are known from the existing art. This problem is
especially acute with liquid enzyme-containing surfactant preparations,
for example liquid washing or cleaning agents. After only a short time
they lose a significant degree of enzymatic, in particular hydrolytic,
and especially proteolytic activity. The surfactant preparation, for
example the washing, cleaning, or disinfecting agent, then no longer
exhibits optimum cleaning performance. One objective in the context of
the development of enzyme-containing surfactant preparations is therefore
to stabilize the contained enzymes and to protect them from denaturing
and/or cleavage or degradation, in particular during storage and/or
during utilization of the surfactant preparation. Hydrolytic enzymes in
particular, and especially proteases, are of interest in this regard.

[0003] Boric acid and boric acid derivatives occupy a prominent position
among the enzyme stabilizers that are effective in surfactant
preparations even at a comparatively low concentration. International
patent application WO 96/21716 A1, for example, discloses that boric acid
derivatives and boronic acid derivatives acting as protease inhibitors
are suitable for stabilizing enzymes in liquid preparations, among them
washing and cleaning agents. A selection of boronic acid derivatives as
stabilizers is disclosed, for example, in international patent
application WO 96/41859 A1. WO 92/19707 A1 and EP 478050 A1 present meta-
and/or para-substituted phenylboronic acids as enzyme stabilizers.
Complexes of boric acids and boric acid derivatives with aromatic
compounds as enzyme stabilizers in liquid detergent compositions are
disclosed in EP 511456 A.

[0004] Boric acids and boric acid derivatives have the disadvantage,
however, that they form undesired secondary products with other
ingredients of a surfactant preparation, in particular washing-,
cleaning-, or disinfecting-agent ingredients, so that they are no longer
available in the relevant agents for the desired cleaning purpose, or in
fact remain behind, for example on the washed item, as a contaminant. In
addition, boric acids or borates are increasingly being regarded as
disadvantageous in environmental terms.

[0005] The underlying object of the present invention is to make available
a liquid surfactant preparation having stabilized hydrolytic enzymes. The
surfactant preparation should preferably contain fewer boron-containing
compounds as enzyme stabilizers.

[0006] The subject matter of the invention is a liquid surfactant
preparation encompassing a hydrolytic enzyme and a component stabilizing
the hydrolytic enzyme, which is characterized in that the component
stabilizing the hydrolytic enzyme encompasses a phthaloylglutamic acid.
Alternatively or in supplementary fashion, the component stabilizing the
hydrolytic enzyme can encompass a phthaloylaspartic acid.

[0007] Furthermore, other desirable features and characteristics of the
present invention will become apparent from the subsequent detailed
description of the invention and the appended claims, taken in
conjunction with the accompanying drawings and this background of the
invention.

[0010] Use of a component that encompasses a phthaloylglutamic acid to
stabilize a hydrolytic enzyme in a liquid surfactant preparation.

[0011] A method, in particular a washing or cleaning method, in which a
hydrolytic enzyme, in particular one that is selected from the group
consisting of protease, amylase, cellulase, glycosidase, hemicellulase,
mannanase, xylanase, xyloglucanase, xanthanase, pectinase,
β-glucosidase, carrageenase, lipase, or mixtures thereof, in
particular a protease, is stabilized in a washing bath by a component
that stabilizes the hydrolytic enzyme and encompasses a phthaloylglutamic
acid.

[0013] The following detailed description of the invention is merely
exemplary in nature and is not intended to limit the invention or the
application and uses of the invention. Furthermore, there is no intention
to be bound by any theory presented in the preceding background of the
invention or the following detailed description of the invention.

[0014] It has been found that a phthaloylglutamic acid and/or a
phthaloylaspartic acid keeps a hydrolytic enzyme, in particular a
protease, advantageously stable in a liquid surfactant preparation, for
example in a liquid washing, cleaning, or disinfecting agent. This opens
up the possibility of using fewer boron-containing compounds as enzyme
stabilizers in liquid surfactant preparations. It is possible in
particular to partly or, by preference, entirely eliminate boric acid as
an enzyme stabilizer in a liquid surfactant preparation, so that the
liquid surfactant preparation can be free of boric acid. In particularly
advantageous embodiments, a surfactant preparation of this kind can
ideally be free of boron.

[0015] In addition, these compounds have the advantage that they already
exert their stabilizing effect at low to very low concentrations. They
moreover possess good water solubility. They can therefore easily be
incorporated or easily utilized in liquid surfactant preparations, in
particular in liquid washing, cleaning, or disinfecting agents or in a
washing bath constituted by such a surfactant preparation. Precipitation
during storage is moreover decreased or entirely avoided.

[0016] The component stabilizing the hydrolytic enzyme encompasses a
phthaloylglutamic acid. This is understood as a substance that is
described by formula (I) below (N-phthaloyl-L-glutamic acid):

##STR00001##

[0017] Alternatively or in supplementary fashion, the component
stabilizing the hydrolytic enzyme can encompass a phthaloylaspartic acid.
This is understood as a substance that is described by formula (II) below
(N-phthaloyl-L-aspartic acid):

##STR00002##

[0018] Also considered a phthaloylglutamic acid or phthaloylaspartic acid
in the context of the invention are derivatives of said compounds. Such
derivatives comprise further chemical modifications; in particular they
can be glycosylated, or they can contain on the phthaloyl residue one or
more methyl, amino, nitro, chloro, fluoro, bromo, hydroxyl, carboxyl,
formyl, ethyl, acetyl, t-butyl, anisyl, benzyl, trifluoroacetyl,
N-hydroxysuccinimide, t-butyloxycarbonyl, benzoyl, 4-methylbenzyl,
thioanizyl, thiocresyl, benzyloxymethyl, 4-nitrophenyl,
benzyloxycarbonyl, 2-nitrobenzoyl, 2-nitrophenylsulfenyl,
4-toluenesulfonyl, pentafluorophenyl, diphenylmethyl,
2-chlorobenzyloxycarbonyl, 2,4,5-trichlorophenyl,
2-bromobenzyloxycarbonyl, 9-fluorenylmethyoxycarbonyl, triphenylmethyl,
2,2,5,7,8-pentamethylchroman-6-sulfonyl residues or groups, or
combinations thereof.

[0019] All compounds that are provided in the context of the present
invention as a component stabilizing the hydrolytic enzyme can be present
in the surfactant preparation in all protonated or deprotonated forms. In
addition, all such compounds, in particular deprotonated forms thereof,
can be associated with cations. Preferred cations in this regard are
divalent cations, in particular calcium ions (Ca2+), magnesium ions
(Mg2+), and zinc ions (Zn2+). Calcium ions (Ca2+) are
particularly preferred.

[0020] The component stabilizing the hydrolytic enzyme can be made up
entirely of the aforesaid compound, so that the component stabilizing the
hydrolytic enzyme is the phthaloylglutamic acid and/or phthaloylaspartic
acid. Alternatively, the component stabilizing the hydrolytic enzyme can
encompass further compounds, so that the phthaloylglutamic acid and/or
phthaloylaspartic acid is part of the component stabilizing the
hydrolytic enzyme.

[0021] The phthaloylglutamic acid and/or phthaloylaspartic acid can
furthermore be present in any possible stereoisomeric form. The
glutamic-acid or aspartic-acid residue in particular can be present in a
D or L configuration, the configuration being identified, in a manner
usual in the art, based on the position of the amino group on the chiral
carbon atom of the glutamic acid or aspartic acid in the Fischer
projection. What is present in the phthaloylglutamic acid or
phthaloylaspartic acid is by preference an L-glutamic acid residue or
L-aspartic acid residue.

[0022] The phthaloylglutamic acid or phthaloylaspartic acid is contained
in the liquid surfactant preparation by preference in a quantity from
0.000001 to 10 wt %, and increasing preferably from 0.00001 to 5 wt %,
from 0.001 to 3 wt %, from 0.01 to 2.5 wt %, from 0.1 to 2.25 wt %, and
from 0.5 to 2 wt %. In the case of combinations of phthaloylglutamic acid
and phthaloylaspartic acid, each compound can be present in the
quantities recited.

[0023] A hydrolytic enzyme is a hydrolase (EC 3.X.X.X) and thus an enzyme
that hydrolytically cleaves esters, ethers, peptides, glycosides, acid
anhydrides, or carbon-carbon bonds in a reversible reaction. The
hydrolytic enzyme therefore catalyzes the hydrolytic cleavage of
substances as defined by: A-B+H2O<->AH+B--OH. Hydrolases form
the third main class in the EC classification of enzymes. The EC (Enzyme
Commission) numbers constitute a numerical classification system for
enzymes. Each EC number is made up of four numbers separated by periods;
the first digit identifies one of the six main enzyme classes, and
hydrolases (EC 3.X.X.X) correspondingly represent the third main class.
Its representatives are proteases, peptidases, nucleases, phosphatases,
glycosidases, and esterases.

[0024] The hydrolytic enzyme is contained in the liquid surfactant
preparation by preference in a quantity from 1×10-8 to 5 weight
percent, based on active protein. The hydrolytic enzyme is contained in
the liquid surfactant preparation preferably from 0.001 to 5 wt %, more
preferably from 0.01 to 5 wt %, even more preferably from 0.05 to 4 wt %,
and particularly preferably from 0.075 to 3.5 wt %. The hydrolytic enzyme
can furthermore be bound covalently or noncovalently to a carrier
substance, and/or embedded into encasing substances, for example in order
to protect it additionally from premature inactivation. The protein
concentration in the surfactant preparation can be determined with the
aid of known methods, for example the BCA method (bicinchoninic acid;
2,2'-biquinolyl-4,4'-dicarboxylic acid) or the biuret method (A. G.
Gornall, C. S. Bardawill and M. M. David, J. Biol. Chem., 177 (1948), pp.
751-766).

[0025] In a further preferred embodiment, a surfactant preparation
according to the present invention is characterized in that the
hydrolytic enzyme is a protease, amylase, cellulase, glycosidase,
hemicellulase, mannanase, xylanase, xyloglucanase, xanthanase, pectinase,
β-glucosidase, carrageenase, or a lipase, or is a mixture that
encompasses at least two of said enzymes. Particularly preferably, the
hydrolytic enzyme is a protease, more preferably a serine protease, more
preferably a subtilase, and very particularly preferably a subtilisin. It
has been found that proteases, in particular such proteases, are
stabilized particularly well by the component stabilizing the hydrolytic
enzyme in a surfactant preparation according to the present invention.
The reason is that the shelf stability of the enzymes, and in particular
also that of proteases, is a general problem especially for washing,
cleaning, or disinfecting agents.

[0026] Examples of proteases are the subtilisins BPN' from Bacillus
amyloliquefaceans and Carlsberg from Bacillus licheniformis, protease
PB92, subtilisins 147 and 309, the protease from Bacillus lentus,
subtilisin DY, and the enzymes (to be classified, however, as subtilases
and no longer as subtilisins in the strict sense) thermitase, proteinase
K, and the proteases TW3 and TW7. Subtilisin Carlsberg is obtainable in
further developed form under the trade name Alcalase® from Novozymes
A/S, Bagsv.ae butted.rd, Denmark. Subtilisins 147 and 309 are marketed by
Novozymes under the trade names Esperase® and Savinase®,
respectively. The protease variants listed under the designation
BLAP® are derived from the protease from Bacillus lentus DSM 5483.
Other usable proteases are, for example, the enzymes obtainable under the
trade names Durazym®, Relase®, Everlase®, Nafizym®,
Natalase®, Kannase®, and Ovozyme® from Novozymes, under the
trade names Purafect®, Purafect® OxP, Purafect® Prime,
Excellase®, and Properase® from Danisco/Genencor, under the trade
name Protosol® from Advanced Biochemicals Ltd., Thane, India, under
the trade name Wuxi® from Wuxi Snyder Bioproducts Ltd., China, under
the trade names Proleather® and Protease P® from Amano
Pharmaceuticals Ltd., Nagoya, Japan, and under the designation Proteinase
K-16 from Kao Corp., Tokyo, Japan. The proteases from Bacillus gibsonii
and Bacillus pumilus, which are disclosed in international patent
applications WO 08/086916 and WO 07/131656, are also used with particular
preference. Further advantageously usable proteases are disclosed in
patent applications WO 91/02792, WO 08/007319, WO 93/18140, WO 01/44452,
GB 1243784, WO 96/34946, WO 02/029024, and WO 03/057246. Further usable
proteases are those that are naturally present in the microorganisms
Stenotrophomonas maltophilia, in particular Stenotrophomonas maltophilia
K279a, Bacillus intermedius, and Bacillus sphaericus.

[0027] Examples of amylases are the α-amylases from Bacillus
licheniformis, from Bacillus amyloliquefaciens, or from Bacillus
stearothermophilus, and in particular the further developments thereof
improved for use in washing or cleaning agents. The enzyme from Bacillus
licheniformus is available from the Novozymes company under the name
Termamyl®, and from Danisco/Genencor under the name Purastar® ST.
Further developed products of this α-amylase are available from
Novozymes under the trade names Duramyl® and Termamyl® ultra,
from Danisco/Genencor under the name Purastar® OxAm, and from Daiwa
Seiko Inc., Tokyo, Japan, as Keistase®. The α-amylase from
Bacillus amyloliquefaciens is marketed by Novozymes under the name
BAN®, and derived variants of the α-amylase from Bacillus
stearothermophilus are marketed, again by Novozymes, under the names
BSG® and Novamyl®. Additionally to be highlighted for this
purpose are the α-amylase from Bacillus sp. A 7-7 (DSM 12368) and
the cyclodextrin-glucanotransferase (CGTase) from Bacillus agaradherens
(DSM 9948). Also usable are the amylolytic enzymes that are disclosed in
international patent applications WO 03/002711, WO 03/054177, and WO
07/079938. Fusion products of all the aforesaid molecules are likewise
usable. The further developments of the α-amylase from Aspergillus
niger and A. oryzae, obtainable from Novozymes under the trade names
Fungamyl®, are also suitable. Further advantageously usable
commercial products are, for example, Amylase-LT® and Stainzyme®
or Stainzyme ultra® or Stainzyme plus®, the latter likewise from
Novozymes. Variants of these enzymes obtainable by point mutations can
also be used according to the present invention.

[0028] Examples of cellulases (endoglucanases, EG) are the fungus-based
cellulase preparation rich in endoglucanase (EG), or its further
developments, offered by the Novozymes company under the trade name
Celluzyme®. The products Endolase® and Carezyme®, likewise
obtainable from the Novozymes company, are based on the 50 kD EG and 43
kD EG, respectively, from Humicola insolens DSM 1800. Further usable
commercial products of this company are Cellusoft®, Renozyme®,
and Celluclean®. Also usable are, for example, cellulases that are
available from the AB Enzymes company, Finland, under the trade names
Ecostone® and Biotouch® and that are based at least in part on
the 20 kD EG from Melanocarpus. Other cellulases of the AB Enzymes
company are Econase® and Ecopulp®. Other suitable cellulases are
from Bacillus sp. CBS 670.93 and CBS 669.83, the one from Bacillus sp.
CBS 670.93 being obtainable from the Danisco/Genencor company under the
trade name Puradax®. Other usable commercial products of the
Danisco/Genencor company are "Genencor detergent cellulase L" and
IndiAge® Neutra.

[0030] Examples of enzymes suitable in this context are obtainable, for
example, under the names Gamanase®, Pektinex AR®, or
Pectaway® from the Novozymes company, under the name Rohapec® B1L
from the AB Enzymes company, and under the name Pyrolase® from
Diversa Corp., San Diego, Calif., USA. The ®-glucanase recovered from
Bacillus subtilis is available under the name Cereflo® from the
Novozymes company. Glycosidases and/or hemicellulases particularly
preferred according to the present invention are mannanases, which are
marketed e.g. under the trade names Mannaway® by Novozymes or
Purabrite® by Danisco/Genencor.

[0031] Examples of lipases or cutinases are the lipases obtainable
originally from Humicola lanuginosa (Thermomyces lanuginosus) and lipases
further developed therefrom, in particular those having the D96L amino
acid exchange. They are marketed, for example, by the Novozymes company
under the trade names Lipolase®, Lipolase® Ultra, LipoPrime®,
Lipozyme®, and Lipex®. A further advantageously usable lipase is
obtainable from the Novozymes company under the trade name
Lipoclean®. The cutinases that were originally isolated from Fusarium
solani pisi and Humicola insolens are moreover usable, for example.
Similarly usable lipases are obtainable from the Amano company under the
designations Lipase CE®, Lipase P®, Lipase B® and Lipase
CES®, Lipase AKG®, Bacillis sp. Lipase®, Lipase AP®,
Lipase M-AP®, and Lipase AML®. The lipases or cutinases from, for
example, the Danisco/Genencor company, whose starting enzymes were
originally isolated from Pseudomonas mendocina and Fusarium solanii, are
usable. To be mentioned as further important commercial products are the
preparations M1 Lipase® and Lipomax® originally marketed by the
Gist-Brocades company (now Danisco/Genencor), and the enzymes marketed by
Meito Sangyo KK, Japan, under the names Lipase MY-30®, Lipase
OF®, and Lipase PL®, as well as the Lumafast® product of the
Danisco/Genencor company.

[0032] The enzymes to be used in the context of the present invention can
originally derive, for example, from microorganisms, e.g. of the genera
Bacillus, Streptomyces, Humicola, or Pseudomonas, and/or can be produced
by suitable microorganisms according to biotechnological methods known
per se, e.g. by means of transgenic expression hosts, for example the
genera Escherichia, Bacillus, or by filamentous fungi. It is emphasized
that this can also involve, in particular, technical enzyme preparations
of the respective enzyme, i.e. accompanying constituents can be present.
The enzymes can therefore be packaged and used together with accompanying
constituents, for example from fermentation, or with further stabilizers.

[0033] Enzyme "stabilization" for purposes of the invention exists when
the presence of the component stabilizing the hydrolytic enzyme causes a
surfactant preparation encompassing hydrolytic enzyme and a component
stabilizing the hydrolytic enzyme (surfactant preparation according to
the present invention) to exhibit after storage a higher enzymatic
activity of the hydrolytic enzyme as compared with a control preparation
that differs from the surfactant preparation according to the present
invention only in that the component stabilizing the hydrolytic enzyme is
absent (control). In this regard, the phthaloylglutamic acid is contained
in the surfactant preparation according to the present invention in a
quantity from 0.5 to 2 wt %. After storage, the surfactant preparation
according to the present invention therefore exhibits higher residual
activity of the hydrolytic enzyme as compared with the control, the
preparation according to the present invention and the control exhibiting
the same initial enzyme activity when storage began, and both
preparations being processed in the same manner, in particular with
regard to storage conditions and the determination of enzyme activity.
Storage occurs, with increasing preference, for at least 24 hours, 48
hours, 72 hours, 5 days, 1 week, 13 days, 3 weeks, or 4 weeks. With
further preference, storage occurs at a temperature of 20° C.,
25° C., or 30° C.

[0034] The enzyme activity can occur in this regard, coordinated with the
respective type of enzyme, in a manner usual in the art. Methods for
determining activity are familiar to one skilled in the art of enzyme
technology, and are routinely utilized by him or her. Methods for
determining protease activity are disclosed, for example, in Tenside,
Vol. 7 (1970), pp. 125-132. Proteolytic activity can furthermore be
determined by way of the release of the para-nitroaniline (pNA)
chromophore from the suc-L-Ala-L-Ala-L-Pro-L-Phe-p-nitroanilide substrate
(suc-AAPF-pNA). The protease cleaves the substrate and releases pNA. The
release of pNA causes an increase in extinction at 410 nm, the time
course of which is an indication of enzymatic activity (see Del Mar et
al., 1979). Measurement is performed at a temperature of 25° C.,
at pH 8.6 and a wavelength of 410 nm. The measurement time is 5 min, with
a measurement interval from 20 s to 60 s. The protease activity is
preferably indicated in CPU (protease units).

[0035] The existence of enzyme stabilization is particularly preferably
ascertained using a protease-containing liquid surfactant preparation
which is stored for 13 days at a temperature of 30° C., and whose
residual proteolytic activity is determined via the release of the
para-nitroaniline (pNA) chromophore from the suc-AAPF-pNA substrate. Very
particularly preferably, the existence of enzyme stabilization in this
regard is ascertained as described in the Example.

[0036] A "surfactant preparation" is to be understood in the context of
the present invention as any type of composition that contains at least
one surfactant. A composition of this kind preferably contains a
surfactant as described below.

[0037] All liquid or flowable administration forms can serve in this
context as liquid surfactant preparations. Preparations that are pourable
and can have viscosities of up to several tens of thousands of mPas are
"flowable" for purposes of the present invention. The viscosity can be
measured with usual standard methods (e.g. Brookfield LVT-II viscosimeter
at 20 rpm and 20° C., spindle 3), and is preferably in the range
from 5 to 10,000 mPas. Preferred agents have viscosities from 10 to 8000
mPas, values between 120 and 3000 mPas being particularly preferred. A
liquid surfactant preparation in the context of the present invention can
therefore also be gel-like or paste-like; it can be present as a
homogeneous solution or suspension, and can, for example, be sprayable or
can be packaged in other usual administration forms.

[0038] A liquid surfactant preparation according to the present invention
can be used as such or after dilution with water, in particular for
cleaning textiles and/or hard surfaces. Such dilution is easily brought
about by diluting a measured quantity of the surfactant preparation in a
further quantity of water at specific weight ratios of surfactant
preparation to water, and optionally shaking that dilution in order to
ensure uniform distribution of the surfactant preparation in water.
Possible weight or volume ratios of the dilutions are from 1:0 surfactant
preparation:water to 1:10,000 or 1:20,000 surfactant preparation:water,
by preference from 1:10 to 1:2000 surfactant preparation:water.

[0039] A "surfactant preparation" for purposes of the present invention
can therefore also be the washing or cleaning bath itself. A "washing or
cleaning bath" is understood as that utilization solution, containing the
washing or cleaning agent, which acts on textiles or fabric (washing
bath) or hard surfaces (cleaning bath) and thereby comes into contact
with stains present on textiles or fabrics or hard surfaces. The washing
or cleaning bath is usually produced when the washing or cleaning
operation begins and the washing or cleaning agent is diluted with water,
for example in a washing machine or in another suitable container.

[0040] In a preferred embodiment, the surfactant preparation is a washing,
cleaning, or disinfecting agent. Included among the washing agents are
all conceivable types of washing agent, in particular washing agents for
textiles, carpets, or natural fibers. They can be provided for manual
and/or also for automatic use. Also included among the washing agents are
washing adjuvants that are dispensed into the actual washing agent in the
context of manual or automatic textile laundering in order to achieve a
further effect. Included among the cleaning agents are all agents, again
occurring in all the aforesaid administration forms, for cleaning and/or
disinfection of hard surfaces, manual and automatic dishwashing agents,
carpet cleaners, scrubbing agents, glass cleaners, toilet deodorizing
cleaners, etc. Lastly, textile pre- and post-treatment agents are on the
one hand those agents with which the laundry item is brought into contact
before actual laundering, for example in order to loosen stubborn stains,
and on the other hand those that, in a step following the actual textile
laundering, impart to the washed item further desirable properties such
as a pleasant feel, freedom from wrinkles, or a low static charge. The
fabric softeners, among others, are categorized among the last-named
agents. Disinfecting agents are, for example, hand disinfecting agents,
surface disinfecting agents, and equipment disinfecting agents, which can
likewise occur in the administration forms mentioned. A disinfecting
agent preferably brings about a germ reduction by a factor of at least
104, i.e. of 10,000 germs originally capable of propagation
(so-called colony-forming units or CFUs), no more than a single one
survives (viruses are not regarded in this context as germs, since they
have no cytoplasm and exhibit no independent metabolism). Preferred
disinfecting agents bring about a germ reduction by a factor of at least
105.

[0041] Anionic, nonionic, zwitterionic, and/or amphoteric surfactants can
be used as surfactant(s). Mixtures of anionic and nonionic surfactants
are preferred in terms of applications engineering. The total surfactant
content of the liquid surfactant preparation is preferably below 60 wt %,
and particularly preferably below 45 wt %, based on the total liquid
surfactant preparation.

[0043] The nonionic surfactants used are by preference alkoxylated,
advantageously ethoxylated, in particular primary alcohols having by
preference 8 to 18 carbon atoms and an average of 1 to 12 mol ethylene
oxide (EO) per mol of alcohol, in which the alcohol residue can be linear
or preferably methyl-branched in the 2-position, or can contain mixed
linear and methyl-branched residues, such as those that are usually
present in oxo alcohol residues. Particularly preferred, however, are
alcohol ethoxylates having linear residues made up of alcohols of natural
origin having 12 to 18 carbon atoms, e.g. from coconut, palm, tallow, or
oleyl alcohol, and an average of 2 to 8 EO per mol of alcohol. The
preferred ethoxylated alcohols include, for example, C12-14 alcohols
with 3 EO, 4 EO or 7 EO, C9-11 alcohol with 7 EO, C13-15
alcohols with 3 EO, 5 EU, 7 EO, or 8 EO, C12-18 alcohols with 3 EO,
5 EO or 7 EO, and mixtures thereof, such as mixtures of C12-14
alcohol with 3 EO and C12-18 alcohol with 7 EO. The degrees of
ethoxylation indicated represent statistical averages, which can
correspond to an integer or a fractional number for a specific product.
Preferred alcohol ethoxylates exhibit a restricted distribution of
homologs (narrow range ethoxylates, NRE). In addition to these nonionic
surfactants, fatty alcohols with more than 12 EO can also be used.
Examples of these are tallow fatty alcohol with 14 EO, 25 EO, 30 EO, or
40 EO. Nonionic surfactants that contain EU and PO groups together in the
molecule are also usable according to the present invention. A mixture of
a (more highly) branched ethoxylated fatty alcohol and an unbranched
ethoxylated fatty alcohol is also suitable, for example a mixture of a
C16-18 fatty alcohol with 7 EO and 2-propylheptanol with 7 EO.
Particularly preferably, the surfactant preparation contains a
C12-18 fatty alcohol with 7 EU or a C13-15 oxoalcohol with 7 EU
as a nonionic surfactant.

[0044] The nonionic surfactant content is preferably 3 to 40 wt %, by
preference 5 to 30 wt %, and in particular 7 to 20 wt %, based in each
case on the total surfactant preparation.

[0045] In addition to the nonionic surfactants, the surfactant preparation
can also contain anionic surfactants. Sulfonates, sulfates, soaps,
alkylphosphates, anionic silicone surfactants, and mixtures thereof are
used by preference as an anionic surfactant.

[0046] Possibilities as surfactants of the sulfonate type are, by
preference, C9-13 alkylbenzenesulfonates, olefinsulfonates, i.e.
mixtures of alkene- and hydroxyalkanesulfonates, and disulfonates, for
example such as those obtained from C12-18 monoolefins having a
terminal or internal double bond, by sulfonation with gaseous sulfur
trioxide and subsequent alkaline or acid hydrolysis of the sulfonation
products. Also suitable are C12-18 alkanesulfonates and the esters
of -sulfo fatty acids (estersulfonates), for example the -sulfonated
methyl esters of hydrogenated coconut, palm kernel, or tallow fatty
acids.

[0047] Preferred alk(en)yl sulfates are the alkali, and in particular
sodium, salts of the sulfuric acid semi-esters of the C12 to
C18 fatty alcohols, for example from coconut fatty alcohol, tallow
fatty alcohol, lauryl, myristyl, cetyl, or stearyl alcohol, or the
C10 to C20 oxo alcohols, and those semi-esters of secondary
alcohols of those chain lengths. For purposes of washing technology, the
C12 to C16 alkyl sulfates and C12 to C15 alkyl
sulfates, as well as C14 to C15 alkyl sulfates, are preferred.
2,3-Alkyl sulfates are also suitable anionic surfactants.

[0048] The sulfuric acid monoesters of straight-chain or branched
C7-21 alcohols ethoxylated with 1 to 6 mol ethylene oxide, such as
2-methyl-branched C9-11 alcohols with an average of 3.5 mol ethylene
oxide (EO), or C12-18 fatty alcohols with 1 to 4 EU, are also
suitable.

[0050] The anionic surfactants, including the soaps, can be present in the
form of their sodium, potassium, magnesium, or ammonium salts. The
anionic surfactants are preferably present in the form of their ammonium
salts. Further preferred counterions for the anionic surfactants are also
the protonated forms of choline, triethylamine, or methylethylamine.

[0051] The concentration of anionic surfactants in a surfactant
preparation can be 1 to 40 wt %, by preference 5 to 30 wt %, and very
particularly preferably 10 to 25 wt %, based in each case on the total
surfactant preparation.

[0052] In a further embodiment, the surfactant preparation is
characterized in that it additionally encompasses at least one further
ingredient that is selected from the group consisting of builder,
nonaqueous solvent, acid, water-soluble salt, thickening agent,
disinfecting ingredient, and combinations thereof.

[0053] The addition of one or more of the further ingredient(s) proves
advantageous because additionally improved cleaning performance and/or
disinfection is thereby achieved. The improved cleaning performance
and/or disinfection is preferably based on a synergistic interaction of
at least two ingredients. A synergy of this kind can be achieved in
particular by way of the combination of the hydrolytic enzyme, by
preference a protease, with one of the builders described below and/or
with one of the nonaqueous solvents described below and/or with one of
the acids described below and/or with one of the water-soluble salts
described below and/or with one of the thickening agents described below
and/or with one of the disinfecting ingredients described below.

[0054] Silicates, aluminum silicates (in particular zeolites), carbonates,
salts of organic di- and polycarboxylic acids, and mixtures of said
substances may be recited, in particular, as builders that can be
contained in the surfactant preparation.

[0055] Organic builders that can be present in the surfactant preparation
are, for example, the polycarboxylic acids usable in the form of the
sodium salts thereof, "polycarboxylic acids" being understood as those
carboxylic acids that carry more than one acid function. These are, for
example, citric acid, adipic acid, succinic acid, glutaric acid, malic
acid, tartaric acid, maleic acid, fumaric acid, sugar acids,
aminocarboxylic acids, nitrilotriacetic acid (NTA), methylglycinediacetic
acid (MGDA), and derivatives thereof, as well as mixtures thereof.
Preferred salts are the salts of the polycarboxylic acids such as citric
acid, adipic acid, succinic acid, glutaric acid, tartaric acid, sugar
acids, and mixtures thereof.

[0056] Polymeric polycarboxylates are additionally suitable as builders.
These are, for example, the alkali-metal salts of polyacrylic acid or of
polymethacrylic acid, for example those having a relative molecular
weight from 600 to 750,000 g/mol.

[0057] Suitable polymers are, in particular, polyacrylates that preferably
have a molecular weight from 1000 to 15,000 g/mol. Of this group in turn,
the short-chain polyacrylates, which have molar masses from 1000 to
10,000 g/mol and particularly preferably from 1000 to 5000 g/mol, may be
preferred because of their superior solubility.

[0058] Also suitable are copolymeric polycarboxylates, in particular those
of acrylic acid with methacrylic acid and of acrylic acid or methacrylic
acid with maleic acid. To improve water solubility, the polymers can also
contain allylsulfonic acids, such as allyloxybenzenesulfonic acid and
methallylsulfonic acid, as monomers.

[0059] It is preferred, however, to use soluble builders, for example
citric acid, or acrylic polymers having a molar mass from 1000 to 5000
g/mol, in the liquid surfactant preparation.

[0060] The molar masses indicated for polymeric polycarboxylates are, for
purposes of this document, weight-average molar masses Mw of the
respective acid form that were determined in principle by means of gel
permeation chromatography (GPC), a UV detector having been used. The
measurement was performed against an external polyacrylic acid standard
that yields realistic molecular weight values because of its structural
affinity with the polymers being investigated. These indications deviate
considerably from the molecular weight indications in which
polystyrenesulfonic acids are used as a standard. The molar masses
measured against polystyrenesulfonic acids are as a rule much higher than
the molar masses indicated in this document.

[0061] Organic builder substances of this kind can be contained, if
desired, in quantities of up to 40 wt %, in particular up to 25 wt %, and
by preference from 1 wt % to 8 wt %. Quantities close to the aforesaid
upper limit are used by preference in pasty or liquid, in particular
water-containing, surfactant preparations.

[0063] In order to establish a desired pH that does not result of itself
from mixture of the other components, the surfactant preparations can
contain system-compatible and environmentally compatible acids, in
particular citric acid, acetic acid, tartaric acid, malic acid, lactic
acid, glycolic acid, succinic acid, glutaric acid, and/or adipic acid,
but also mineral acids, in particular sulfuric acid, or bases, in
particular ammonium hydroxides or alkali hydroxides. pH regulators of
this kind are contained in the surfactant preparations in quantities by
preference not above 20 wt %, in particular from 1.2 wt % to 17 wt %.

[0064] A surfactant preparation for purposes of the invention can
furthermore contain one or more water-soluble salts, which serve e.g. to
adjust viscosity. These can be inorganic and/or organic salts. Usable
inorganic salts are selected in this context by preference from the group
encompassing colorless water-soluble halides, sulfates, sulfites,
carbonates, hydrogencarbonates, nitrates, nitrites, phosphates, and/or
oxides of the alkali metals, of the alkaline earth metals, of aluminum,
and/or of the transition metals; ammonium salts are also usable. Halides
and sulfates of the alkali metals are particularly preferred in this
context; the inorganic salt is therefore preferably selected from the
group encompassing sodium chloride, potassium chloride, sodium sulfate,
potassium sulfate, and mixtures thereof. Usable organic salts are, for
example, colorless water-soluble alkali-metal, alkaline-earth-metal,
ammonium, aluminum, and/or transition-metal salts of carboxylic acids.
The salts are by preference selected from the group encompassing formate,
acetate, propionate, citrate, malate, tartrate, succinate, malonate,
oxalate, lactate, and mixtures thereof.

[0065] For thickening, a surfactant preparation according to the present
invention can contain one or more thickening agents. The thickening agent
is preferably selected from the group encompassing xanthan, guar,
carrageenan, agar-agar, gellan, pectin, locust bean flour, and mixtures
thereof. These compounds are effective thickening agents even in the
presence of inorganic salts. In a particularly preferred embodiment, the
surfactant preparation contains xanthan as a thickening agent, since
xanthan thickens effectively even in the presence of high salt
concentrations and prevents macroscopic separation of the continuous
phase. In addition, the thickening agent stabilizes the continuous,
surfactant-poor phase and prevents macroscopic phase separation.

[0066] Alternatively or in supplementary fashion, (meth)acrylic acid
(co)polymers can also be used as thickening agents. Suitable acrylic and
methacrylic (co)polymers encompass, for example, the
high-molecular-weight homopolymers of acrylic acid crosslinked with a
polyalkenyl polyether, in particular an allyl ether, of sucrose,
pentaerythritol, or propylene (INCI name, according to "International
Dictionary of Cosmetic Ingredients" of the Cosmetic, Toiletry and
Fragrance Association (CFTA): Carbomer), which are also referred to as
carboxyvinyl polymers. Polyacrylic acids of this kind are obtainable,
inter alia, under the trade names Polygel® and Carbopol®. Also
suitable, for example, are the following acrylic acid copolymers: (i)
copolymers of two or more monomers from the group of acrylic acid,
methacrylic acid, and their simple esters, formed by preference with
C1-4 alkanols (INCI: Acrylates Copolymer), which are obtainable, for
example, under the trade names Aculyn®, Acusol®, or Tego®
Polymer, (ii) crosslinked high-molecular-weight acrylic acid copolymers,
included among which are, for example, the copolymers, crosslinked with
an allyl ether of sucrose or of pentaerythritol, of C10-30 alkyl
acrylates with one or more monomers from the group of acrylic acid,
methacrylic acid, and their simple esters formed preferably with
C1-4 alkanols (INCI: Acrylates/C10-30 Alkyl Acrylate
Crosspolymer), and which are obtainable, for example, under the trade
name Carbopol®. Further suitable polymers are (meth)acrylic acid
(co)polymers of the Sokalan® type.

[0067] It may be preferred for the surfactant preparation according to the
present invention to contain a (meth)acrylic acid (co)polymer in
combination with a further thickening agent, by preference xanthan. The
surfactant preparation can contain 0.05 to 1.5 wt %, and by preference
0.1 to 1 wt % thickening agent, based in each case on the total
surfactant preparation. The quantity of thickening agent used depends
here on the nature of the thickening agent and the desired degree of
thickening.

[0068] A "disinfecting ingredient" is understood in particular as
ingredients that possess an antimicrobial or antiviral effect, i.e. that
kill germs. The germ-killing effect depends in this context on the
concentration of the disinfecting ingredient in the surfactant
preparation; the germ-killing effect decreases as the concentration of
the disinfecting ingredient decreases, or as the dilution of the
surfactant preparation increases.

[0069] A preferred disinfecting ingredient is ethanol or propanol. These
monovalent alcohols are often used in disinfecting agents, and also in
cleaning agents in general, because of their solvent properties and their
germ-killing effect. The term "propanol" here encompasses both 1-propanol
(n-propanol) and 2-propanol (isopropanol). Ethanol and/or propanol is
contained in the surfactant preparation, for example, in a total quantity
from 10 to 65 wt %, by preference 25 to 55 wt %. A further preferred
disinfecting ingredient is tea tree oil. This is the essential oil of the
Australian tea tree (Melaleuca alternifolia), an evergreen shrub of the
Melaleuca genus native to New South Wales and Queensland, and of further
tea tree species of various genera (e.g. Baeckea, Kunzea, and
Leptospermum) in the Myrtaceae family. Tea tree oil is obtained by steam
distillation from the leaves and twigs of these trees, and is a mixture
of approx. 100 substances; among the principal constituents are
(+)-terpinen-4-ol, α-terpinene, terpinolene, terpineol, pinene,
myrcene, phellandrene, p-cymene, limonene, and 1,8-cineole. Tea tree oil
is contained in the virucidal treatment solution, for example, in a
quantity from 0.05 to 10 wt %, by preference 0.1 to 5.0 wt %. A further
preferred disinfecting ingredient is lactic acid. Lactic acid, or
2-hydroxypropionic acid, is a fermentation product that is generated by a
variety of microorganisms. It has mild antibiotic activity. Lactic acid
is contained in the surfactant preparation, for example, in quantities of
up to 10 wt %, by preference 0.2 to 5.0 wt %.

[0071] Liquid surfactant preparations according to the present invention
in the form of solutions containing usual solvents are manufactured as a
rule by simply mixing the ingredients, which can be placed into an
automatic mixer as substance or as solution.

[0072] Surfactant preparations according to the present invention can
contain only the hydrolytic enzyme as described. Alternatively, they can
also contain further hydrolytic enzymes or other enzymes at a
concentration useful for the effectiveness of the surfactant preparation.
A further subject of the invention is thus represented by surfactant
preparations that additionally encompass one or more further enzymes, all
enzymes established in the existing art for these purposes being usable
in principle. All enzymes that can display a catalytic activity in a
surfactant preparation according to the present invention are preferably
usable as further enzymes, in particular a protease, amylase, cellulase,
hemicellulase, mannanase, tannase, xylanase, xanthanase, xyloglucanase,
β-glucosidase, pectinase, carrageenase, perhydrolase, oxidase,
oxidoreductase, or a lipase, as well as mixtures thereof. Further enzymes
are contained in the surfactant preparation advantageously in a
respective total quantity from 1×10-8 to 5 weight percent,
based on active protein. Each enzyme is contained in surfactant
preparations according to the present invention preferably from 0.0001 to
1% and more preferably from 0.0005 to 0.5%, 0.001 to 0.1%, and
particularly preferably from 0.001 to 0.06 wt %, based on active protein.
Particularly preferably, the enzymes exhibit synergistic cleaning
performance results with respect to specific stains or spots, i.e. the
enzymes contained in the surfactant preparation mutually assist one
another in terms of their cleaning performance. Very particularly
preferably, a synergy of this kind exists between a contained protease
and a further enzyme of an agent according to the present invention,
thereamong in particular between the protease and a lipase and/or an
amylase and/or a mannanase and/or a cellulase and/or a pectinase.
Synergistic effects can occur not only between various enzymes, but also
between one or more enzymes and further ingredients of the surfactant
preparation according to the present invention.

[0073] In a surfactant preparation according to the present invention, the
component stabilizing the hydrolytic enzyme can moreover encompass at
least one further enzyme stabilizer. A further enzyme stabilizer of this
kind is or encompasses, for example, a polyol, in particular glycerol,
1,2-ethylene glycol or propylene glycol, an antioxidant, glyceric acid,
calcium ions or calcium compounds, lactate, or a lactate derivative. It
can also involve one or more of those enzyme-stabilizing compounds which
are disclosed in the international patent applications WO 07/113241 A1 or
WO 02/008398 A1. The interaction of phthaloylglutamic acid and the
further enzyme stabilizer preferably results in synergistic enzyme
stabilization. This is understood to be better enzyme stabilization by
the combination of the two compounds as compared with enzyme
stabilization by each one of said compounds alone, and also as compared
with the sum of the individual performance results of the two compounds
in terms of enzyme stabilization. A combination of corresponding
compounds as the component stabilizing the hydrolytic enzyme thus makes
it possible, for example, to use the stabilizers in surfactant
preparations according to the present invention in lower concentrations
in total. It is further possible to achieve improved enzyme stabilization
with a component of this kind stabilizing the hydrolytic enzyme. In this
regard, the further enzyme stabilizer does not necessarily need to be a
boron-free stabilizer, since it is also possible, because of the
interaction of the two compounds, to use a smaller quantity of a
boron-containing compound in a surfactant preparation. For example, it is
also possible in this regard to use a phenylboronic acid derivative
having the structural formula

##STR00003##

in which R denotes hydrogen, a hydroxyl group, a C1 to C6 alkyl group, a
substituted C1 to C6 alkyl group, a C1 to C6 alkenyl group, or a
substituted C1 to C6 alkenyl group, by preference 4-formylphenylboronic
acid (4-FPBA), as a further enzyme stabilizer.

[0074] The further enzyme stabilizer is present in the surfactant
preparation by preference in a concentration from 0.000001 to 10 wt %,
and increasingly preferably from 0.00001 to 5 wt %, from 0.0001 to 2.5 wt
%, from 0.001 to 2 wt %, from 0.01 to 1.5 wt %, and from 0.1 to 1 wt %.

[0075] A further subject of the invention is the use of a component that
encompasses a phthaloylglutamic acid to stabilize a hydrolytic enzyme in
a liquid surfactant preparation.

[0076] Alternatively or in supplementary fashion, the component
stabilizing the hydrolytic enzyme can encompass a phthaloylaspartic acid.

[0077] The reason is that, as set forth above, an advantageous
stabilization of the hydrolytic enzyme in a liquid surfactant preparation
is achieved by this/these component(s). The hydrolytic enzyme is by
preference a protease.

[0078] All facts, subjects, and embodiments that are described for
surfactant preparations according to the present invention are also
applicable to this subject of the invention. Reference is therefore made
at this junction expressly to the disclosure at the corresponding
location, with the instruction that said disclosure also applies to the
present use according to the present invention.

[0079] A further subject of the invention is a method in which a
hydrolytic enzyme is stabilized in a washing bath by a component that
stabilizes the hydrolytic enzyme and encompasses a phthaloylglutamic
acid.

[0080] Alternatively or in supplementary fashion, the component
stabilizing the hydrolytic enzyme can encompass a phthaloylaspartic acid.

[0081] The reason is that, as set forth above, an advantageous
stabilization of the hydrolytic enzyme in a liquid surfactant preparation
is achieved by this/these component(s). The hydrolytic enzyme is
consequently also stabilized in the corresponding washing or cleaning
bath whose basis is the liquid surfactant preparation. The method is
preferably a washing, cleaning, or disinfecting method. Particularly
preferably, a surfactant preparation as described above is utilized in
such a method. By preference, the hydrolytic enzyme is selected from the
group consisting of protease, amylase, cellulase, glycosidase,
hemicellulase, mannanase, xylanase, xyloglucanase, xanthanase, pectinase,
β-glucosidase, carrageenase, lipase, or mixtures thereof.
Particularly preferably, the hydrolytic enzyme is a protease.

[0082] A method according to the present invention preferably occurs in a
temperature range between 10° C. and 60° C., in particular
between 10° C. and 50° C., between 10° C. and
40° C., between 10° C. and 30° C., and particularly
preferably between 15° C. and 30° C. Thermally stable
hydrolytic enzymes could also be used in methods according to the present
invention even at temperatures higher than 60° C., for example up
to 70° C. or 75° C. The pH at which a method according to
the present invention is advantageously carried out can be dependent on
the object to be treated. For example, a surfactant preparation that is
based on a cleaning agent for toilets advantageously has an acid pH, for
example a pH between pH 2 and pH 5. A surfactant preparation that is
based on a textile washing agent or a cleaning agent for other hard
surfaces advantageously has a slightly acid, neutral, or alkaline pH, for
example a pH between pH 6 and pH 11 or between pH 7 and pH 10. A
surfactant preparation that is based on a hand dishwashing agent has, for
example, a pH of between pH 6.5 and pH 8. It is consequently advantageous
also to carry out a method according to the present invention at these
respective pH values.

[0083] All facts, subjects, and embodiments that are described for
surfactant preparations according to the present invention are also
applicable to this subject of the invention. Reference is therefore made
at this junction expressly to the disclosure at the corresponding
location, with the instruction that said disclosure also applies to
methods according to the present invention.

[0084] A further subject of the invention is a liquid enzyme preparation
encompassing a hydrolytic enzyme and a component stabilizing the
hydrolytic enzyme, which is characterized in that the component
stabilizing the hydrolytic enzyme encompasses a phthaloylglutamic acid.

[0085] Alternatively or in supplementary fashion, the component
stabilizing the hydrolytic enzyme can encompass a phthaloylaspartic acid.

[0086] It has been determined that a component stabilizing the hydrolytic
enzyme as described above also stabilizes a hydrolytic enzyme in a liquid
preparation that encompasses no surfactant. With such a component it is
consequently possible also to stabilize hydrolytic enzymes in a culture
supernatant of a fermentation, during the processing of a culture
supernatant of a fermentation, or in a liquid enzyme preparation. By
preference, the phthaloylglutamic acid or phthaloylaspartic acid is
contained in the preparation in a quantity from 0.000001 to 10 wt %,
and/or the hydrolytic enzyme is contained in a quantity from
1×10-8 to 5 wt %, based on active protein. In the case of
combinations of phthaloylglutamic acid and phthaloylaspartic acid, each
compound can be present in the quantity recited. Also preferably, the
hydrolytic enzyme is a protease. All further facts, subjects, and
embodiments that are not applicable exclusively to surfactant
preparations according to the present invention are consequently also
applicable to this subject of the invention. Reference is therefore made
at this junction expressly to the disclosure at the corresponding
location, with the instruction that said disclosure also applies to
liquid enzyme preparations according to the present invention.

[0087] Example: Stabilizing a protease in a liquid washing agent according
to the present invention

[0089] The phthaloylglutamic acid N-phthaloyl-L-glutamic acid (Fluka) was
incorporated into this formulation as the component stabilizing the
hydrolytic enzyme, as indicated below (see Table 1, indications in this
regard in wt %). Comparison formulations that contained either boric acid
as an enzyme stabilizer, or no enzyme stabilizer, served as controls. The
protease used was variant F49 of the protease from Bacillus lentus in
accordance with WO 95/23221 (quantity used: 1 wt % active substance).

[0090] Storage occurred in airtight sealed vessels at 30° C. over
time periods of various lengths as indicated in Table 1. After storage,
the respective residual proteolytic activity was determined by way of the
release of the para-nitroaniline @NA) chromophore from the
suc-L-Ala-L-Ala-L-Pro-L-Phe-p-nitroanilide substrate (suc-AAPF-pNA). The
protease cleaves the substrate and releases pNA. The release of pNA
causes an increase in extinction at 410 nm, the time course of which is
an indication of enzymatic activity (see Del Mar et al., 1979).
Measurement occurred at a temperature of 25° C., at pH 8.6 and at
a wavelength of 410 nm. The measurement time was 5 min, with a
measurement interval from 20 s to 60 s. The proteolytic activity values
obtained are indicated in Table 1 below, based on an initial activity of
100% when storage began.

[0091] It is evident that a component according to the present invention
that stabilizes the hydrolytic enzyme produces an improvement in enzyme
stability as compared with the control having no enzyme stabilizer. It
can consequently be used in order to partly or entirely eliminate boric
acid or boron-containing compounds as an enzyme stabilizer in a liquid
surfactant preparation.

[0092] While at least one exemplary embodiment has been presented in the
foregoing detailed description of the invention, it should be appreciated
that a vast number of variations exist. It should also be appreciated
that the exemplary embodiment or exemplary embodiments are only examples,
and are not intended to limit the scope, applicability, or configuration
of the invention in any way. Rather, the foregoing detailed description
will provide those skilled in the art with a convenient road map for
implementing an exemplary embodiment of the invention, it being
understood that various changes may be made in the function and
arrangement of elements described in an exemplary embodiment without
departing from the scope of the invention as set forth in the appended
claims and their legal equivalents.

Patent applications by Hendrik Hellmuth, Duesseldorf DE

Patent applications by Karl-Heinz Maurer, Erkrath DE

Patent applications by Marion Merkel, Koeln DE

Patent applications by Petra Siegert, Haan DE

Patent applications by Timothy O'Connell, Duesseldorf DE

Patent applications by Henkel AG & Co., KGaA

Patent applications in class Stablizing an enzyme by forming a mixture, an adduct or a composition, or formation of an adduct or enzyme conjugate

Patent applications in all subclasses Stablizing an enzyme by forming a mixture, an adduct or a composition, or formation of an adduct or enzyme conjugate